Objective To investigate the effect of antisense oligodeoxynueleotide targeting sur-vivin (survivin ASODN) on hilar cholangioearcinoma cell line FRH-0201 depressing the expression of survivin. Methods Survivin ASODN was transfected into hilar eholangiocarcinoma cell line FRH-0201by liposome. Morphologieal changes were observed by inverted phase contrast microscope. RT-PCR and Western blot methods were performed to detect the expressions of survivin mRNA and protein re-spectively. The changes in cell apoptosis were detected by flow cytometry. Results The expression of survivin mRNA and protein was significantly decreased in survivin ASODN group than that in the con-trol group(mRNA: 0.51-t-0. 03 vs 0. 82-t-0.02,P%0. 05～protein: 1.82-t-0.16 vs 3. 08--t-_0. 27, P-Q 0.05). The morphologieal apoptotic changes were observed and the apoptosis rate was increased (11.50+1.49% vs 0.39-+-0.08～, P%0.05). Conclusion Survivin AS()DN can induce hilar cholan-gioeareinoma cell line FRH-0201 into apoptosis by decreasing the expression of survivin.

OBJECTIVE: To discuss the application of humanized mice model in drug metabolism and toxicity. METHODS: By sorting foreign scientific literature, the history of the development of humanized mice model and its application in drug metabolizing enzymes and toxicity studies were reviewed and summarized to provide support for early drug development. RESULTS and CONCLUSION: The humanized mice model could reduce interspecies differences and has been increasingly used in drug metabolism and safety evaluation in foreign countries. Transgenic mice can be applied to studying the metabolism of drugs metabolized by certain enzyme. In the chimeric mice body, the disposal of drugs could be more systematic and comprehensive mirrored, predicting this model has enormous potential in drug discovery and development.

To investigate the relationship between transcription factor and the change of protein expression levels in heart failure. Methods Bioinformatic method was used to analyze the data of binding-sites on the 5 ' flaking regions of four genes whose mRNA level changed in failing heart from three databases about nucleic acid-EMBL, transcriptional regulation factor-TRANSFAC and protein-SWISS-PORT.The expression level of selected transcription factor was determined by immunohischemical method.Results Nine transcription factors were inferred to influence the proteins' levels in occurrence and development of heart failure.Serum response factor (SRF) was selected from the nine factors and assayed. The results showed that there was a higher level of SRF in healthy group than in chronic heart failure (CHF), and the level was associated with the degree of CHF. It was also found that there was a relative higher level of SRF in the acute myocardial infarction (AMI) than that in CHF, but which was lower than the healthy. Conclusion It showed that SRF had a quantitative change in the development of heart failure, and suggested SRF might play an important regulative role in heart failure. The expression changes of proteins related to myocardial function might be regulated by the quantitative change of transcription factor(s).

Objective To explore the performance of the novel nitric oxide(NO) electrochemical microsensor based on Au nano-particles(nano-Au) modified glass-fiber .Methods With columnar glass-fiber as the substrate material ,the in-situ chemical seed-growth technology was adopted to fabricate one kind of electrochemical microsensor based on gold nanoparticles pole electrodes and the amperometric response method was used to investigate the various properties of the NO microsensor .Results This microsensor had high sensitivity response to NO ,its linear range was 7 .2 nmol/L -11 .7μmol/L(r=0 .998) with a detection limit of 3 .6 nmol/L .Conclusion A novel electrochemical microsensor based on nano-Au pole electrode was fabricated .The NO microsensor posseses high sensitivity ,wide linear range ,good reproducibility and excellent anti-interference ability .

· AIM: To investigate the efficacy and safety of intravitreal injection of plasmin, hyaluronidase, or the combination of the two in inducing posterior vitreous detachment (PVD).· METHODS: 15 mini-type pigs were assigned to three groups (Group A, B and C), 5 in each group. One eye of each pig was intravitreally injected with the studying agent,and the fellow eye was used as control. Group A received a vitreous injection of hyaluronidase 50U (0.1mL); group B received plasmin 0.5U (0.1mL); group C received plasmin 0.5U (0.05mL) combined with hyaluronidase 50U (0.05mL). The fellow eyes in each group were injected with 0.1mL balanced salt solution (BSS). All the pigs were examined with slit-lamp biomicroscope, direct and indirect ophthalmoscope, B-scan and electroretinograph (ERG). After 7 days, the animals were killed and the eyes were enucleated and examined with light microscope, transmission electron microscope and scanning electron microscope.· RESULTS: B-scan examination showed that one eye of Group A and two eyes of Group B had partial PVD at 1st day after injection and one eye of Group C at 1 hour after injection. On the 7th day, B-scan, light microscopy and scanning electron microscopy demonstrated that all the eyes of Group A and Group B had partial PVD, while none of the control eyes had PVD. Rank sum test for scanning electron microscopy results of all the groups showed P ＜0.005.Furthermore, the comparisons between every two groups were made. The results of analyses were as follows: P＞0.05 between the drug injected eyes of Group A and Group B, P＜0.05 between Group B and Group C, Group A and Group C.The b-wave and a-wave amplitudes of ERG showed no significant difference either between preinjection and postinjection in all groups or between the drug injected eyes and the control eyes in each group. Light microscopy and transmission electron microscopy revealed no damage to the retinal structure.· CONCLUSION: Intravitreal injection of hyaluronidase 50U or plasmin 0.5U or their combination can produce PVD effectively and quickly without retinal functional or structural toxicity. The combination of the two proteases was proved to be synergetic.

Animals ; Cytokines ; blood ; Endotoxemia ; blood ; therapy ; Endotoxins ; blood ; toxicity ; Enzyme-Linked Immunosorbent Assay ; Hemofiltration ; Interleukin-1 ; blood ; Interleukin-10 ; blood ; Models, Animal ; Swine ; Time Factors ; Tumor Necrosis Factor-alpha ; analysis

To breed a new yam cultivar of Dioscorea alata, the different and excellent germplasm resources were investigated within artificially cultivated population and some superior individuals, with a higher yield and medicinal properties, were selected. Considering results of the yield and medicinal properties during 2006-2013 cropping season, strains and lines were established and selected. As a result, the yield of the new developed cultivar (Wenshanyao No. 1, WSY01-1) reached 2217. 0 kg per 667 m2 (fresh weight) and 348.3 kg per 667 m2 (dry weight), and increased 23.8% and 23.9% comparing with control cultivars (landraces). Comparing with control cultivars, the level of polysaccharide, allantoin, and dioscin increased 36.9%, 48.3%, 20.9%, and reached 12.2%, 1.30%, 579.7 µg · g(-1), respectively. This result showed that the systematic selection method can significantly improve yield and medicinal properties of D. alata, and the developed " Wenshanyao No. 1" exhibits wide spreading prospects.
Allantoin ; analysis ; Breeding ; Dioscorea ; chemistry ; genetics ; growth & development ; Diosgenin ; analogs & derivatives ; analysis ; Polysaccharides ; analysis

This study aims to establish a new method for quality evaluation of Gentianae Macrophyllae Radix by simultaneous determination of five iridoids (loganic acid, 6'-O-beta-D-glucopyranosylgentiopicroside, swertiamarin, gentiopicroside, sweroside), and to detect five iridoids in the root of eight species (Gentiana macrophylla, G. straminea, G. crassicaulis, G. dahurica, G. robusta, G. waltonii, G. lhassica, and G. tibetica). The separation was carried out on a Shiseido SPOLAR C18 (4.6 mm x 250 mm, 5 microm) column eluted with mobile phase of water containing 0.04% formic acid (A) and acetonitrile (B) in a gradient program. The flow rate was 0.8 mL x min(-1). The detect wavelength was set at 240 nm. The column temperature was kept at 30 degrees C. The volume of injection was 5 microL. The five iridoids were well separated with ideal linear correlations. The average recoveries were 97.35% - 106.23%. All the five iridoids were detected in the root of eight species. The contents of same species changed in a somewhat wider range. The contents in root of G. dahurica were lower than that in other species.
China ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Gentianella ; chemistry ; Iridoid Glycosides ; analysis

In order to breed and spread a new cultivar of Curcuma wenyujin, the C. wenyujin germplasm resources were investigated in authentic regions. Better varieties were chosen by comparing the yield, economic characters and quality differences between different cultivars. The results showed that the character of new selected cultivar was stable, the yield of zedoary, turmeric and curcuma was reached 313.7, 177.9, 91.2 kg per 667 m2, respectively, it increased 11.6%, 10.2%, 14.2% comparing with farmer varieties. The volatile oil contents in zedoary and turmeric was 4.0%, 3.0%, respectively. The target ingredients (germacrone) content was stable. It is demonstrated that the new cultivar "Wenyujin No. 1" has value for extension at authentic regions.
Breeding ; China ; Curcuma ; chemistry ; growth & development ; Oils, Volatile ; analysis ; Plant Extracts ; analysis

Objective To explore the expression and its implication of angiogenesis and invasiveness related factor in primary and recurrent glioma. Methods Expressions of vascular endothelial growth factor (VEGF), matrix metalloproteinase-2 (MMP-2) and MMP-9 were detected by immunohistochemical technique. The morphological characteristics of ultrastructure of glioma were observed by transmission electron microscope (TEM). Results The expressions of VEGF, MMP-2 and MMP-9 varied in different grades of primary glioma. With the elevation of the malignant degree of the primary glioma, positive staining rates of VEGF, MMP-2 and MMP-9 increased significantly. The expression of VEGF correlated with both MMP-2 and MMP-9 expression. Compared with the primary glioma, the immunoreactivities of VEGF, MMP-2 and MMP-9 in recurrent glioma increased, especially in those with more severe malignancy. Under transmission electron microscope, endothelial cells markedly proliferated and protruded from the deficiency of basemembrane, concomitantly with edema of the extracapillary gap, plasma extravasation as well as some small worm-eaten caverns in the basemembrane.Conclusion VEGF, MMP-2 and MMP-9 play important roles in glioma angiogenesis and invasiveness.Inhibition of their expressions may be a useful therapy to glioma.